DESCRIPTION: We propose to develop an integrated diagnostics system that provides ultrasensitive, low cost and very rapid food quality diagnostics. This new methodology will be implemented initially as tests for contamination by pathogenic strains of E. coli. We will apply Multi Photon Detection (MPD) technology to food diagnostics to accomplish the following: A fast and very sensitive MPD enhanced quantitative PCR; An ultra fast and very sensitive MPD enhanced direct detection (i.e., non- amplification based) DNA assay for food contaminants; and Validation of this new methodology by comparison of MPD enhanced assays with conventional microbiological diagnostic methods. The main goal is to develop methodologies that translate the superior sensitivity of MPD into speed and reliability of food tests. These new methods are generic and can be used for all important food contaminants. Ultra-sensitive MPD already has been used to quantitate sub-zeptomole amounts (less than 10-21 M) of biological macromolecules such as DNA, RNA and proteins. Our prototype MPD Imager has enabled the development of super-sensitive tests using both immuno techniques and DNA probes. As an example, we developed a rapid diagnostic assay that permits reliable quantitation of a few copies of DNA in a few milliliters of sample. PROPOSED COMMERCIAL APPLICATION: If successful, this project will lead to the development of a good quality diagnostics system which provides rapid, high sensitivity and economical testing for major food borne pathogens, This system will be designed to operate in a food processing plant environment and will offer the ability to detect as little as a single viable pathogenic organism in a food sample in less that six hours. This unique capability will enable the BioTraces food pathogen system to capture a significant share of the market for in-plant testing for food borne pathogens, which now totals over 25 million tests per year with double digit growth rates. [unreadable]